Life Technologies

4 Life Technologies ™ | Parkinson's cell models Research summary Collaboration summary In 2012, we partnered with the Parkinson's Institute and Clinical Center in Sunnyvale, California, to develop PD model systems using fibroblasts from donor skin biopsies that were collected at the Institute. We used these fibroblasts to generate a fully characterized set of induced pluripotent stem cells (iPSCs). Read Part 1 - Generation of iPSCs. From these iPSCs, we generated and characterized neural stem cells (NSCs), which were shown to have the potential to be further differentiated to dopaminergic neurons, astrocytes, or glia. Read Part 2 - Generation of NSCs. Using these NSCs, we developed a set of cellular assays that allowed us to monitor PD-relevant phenotypes. In this third part of our series, we describe the use of GeneArt ® Precision TALs technology [2] to modify disease-relevant genes in these cell lines. The resulting cell lines will allow us to study the contribution of specific mutations in PD-related genes to the disease phenotype. To study the impact that α-synuclein levels have on disease-related phenotypes, we first generated cell lines in which either one or both copies of SNCA, the gene coding for α-synuclein, were deleted (knocked out) from the iPSCs generated from a 62-year-old female donor with Multiple System Atrophy (MSA). MSA is a Parkinsonian disorder with no known genetic risk factors and is characterized by pronounced α-synuclein deposits called Lewy bodies [3]. Using GeneArt ® Precision TALs technology, the SNCA gene was knocked out by a non-homologous end joining (NHEJ) repair mechanism. The two resulting cell lines will allow for future studies on the What is Parkinson's disease? Parkinson's disease (PD) is a progressive neurodegenerative disorder that affects 1% of people over age 60 and more than 5 million people worldwide [1]. PD results primarily from the selective loss of dopaminergic neurons in the substantia nigra pars compacta region of the brain. The loss of these neurons first affects movement, but then later affects cognitive function, with late-stage disease often being accompanied by dementia. The absence of physiologically relevant cellular models for PD represents a major bottleneck for PD research, and such models are urgently needed to accelerate the discovery of disease mechanisms or drug targets, or to identify and develop therapeutics. To meet this need, tissues derived from disease-affected donor individuals have become an attractive tool for modeling disease in vitro. Dopamine system Frontal lobe Ventral tegmental area Striatum Substantia nigra CULTURE ENGINEERING DIFFERENTIATION CHARACTERIZATION

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